Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H

Y. Inaguma, Y. Akahori, Y. Murayama, K. Shiraishi, S. Tsuzuki-Iba, A. Endoh, J. Tsujikawa, A. Demachi-Okamura, K. Hiramatsu, H. Saji, Y. Yamamoto, Naoki Yamamoto, Y. Nishimura, T. Takahashi, K. Kuzushima, N. Emi, Yoshiki Akatsuka

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The genetic transfer of T-cell receptors (TCRs) directed toward target antigens into T lymphocytes has been used to generate antitumor T cells efficiently without the need for the in vitro induction and expansion of T cells with cognate specificity. Alternatively, T cells have been gene-modified with a TCR-like antibody or chimeric antigen receptor (CAR). We show that immunization of HLA-A2 transgenic mice with tetramerized recombinant HLA-A2 incorporating HA-1 H minor histocompatibility antigen (mHag) peptides and β2- microglobulin (HA-1 H/HLA-A2) generate highly specific antibodies. One single-chain variable region moiety (scFv) antibody, #131, demonstrated high affinity (K D =14.9 nM) for the HA-1 H/HLA-A2 complex. Primary human T cells transduced with #131 scFV coupled to CD28 transmembrane and CD3ζ domains were stained with HA-1 H/HLA-A2 tetramers slightly more intensely than a cytotoxic T lymphocyte (CTL) clone specific for endogenously HLA-A2-and HA-1 H-positive cells. Although #131 scFv CAR-T cells required >100-fold higher antigen density to exert cytotoxicity compared with the cognate CTL clone, they could produce inflammatory cytokines against cells expressing HLA-A2 and HA-1 H transgenes. These data implicate that T cells with high-affinity antigen receptors reduce the ability to lyse targets with low-density peptide/MHC complexes (∼100 per cell), while they could respond at cytokine production level.

Original languageEnglish
Pages (from-to)575-584
Number of pages10
JournalGene Therapy
Volume21
Issue number6
DOIs
Publication statusPublished - 01-01-2014

Fingerprint

Minor Histocompatibility Antigens
HLA-A2 Antigen
T-Cell Antigen Receptor
Antibodies
T-Lymphocytes
Antigen Receptors
Cytotoxic T-Lymphocytes
Clone Cells
Cytokines
Antigens
Single-Chain Antibodies
Transgenes
Transgenic Mice
Immunization
Peptides

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Inaguma, Y., Akahori, Y., Murayama, Y., Shiraishi, K., Tsuzuki-Iba, S., Endoh, A., ... Akatsuka, Y. (2014). Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. Gene Therapy, 21(6), 575-584. https://doi.org/10.1038/gt.2014.30
Inaguma, Y. ; Akahori, Y. ; Murayama, Y. ; Shiraishi, K. ; Tsuzuki-Iba, S. ; Endoh, A. ; Tsujikawa, J. ; Demachi-Okamura, A. ; Hiramatsu, K. ; Saji, H. ; Yamamoto, Y. ; Yamamoto, Naoki ; Nishimura, Y. ; Takahashi, T. ; Kuzushima, K. ; Emi, N. ; Akatsuka, Yoshiki. / Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. In: Gene Therapy. 2014 ; Vol. 21, No. 6. pp. 575-584.
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Inaguma, Y, Akahori, Y, Murayama, Y, Shiraishi, K, Tsuzuki-Iba, S, Endoh, A, Tsujikawa, J, Demachi-Okamura, A, Hiramatsu, K, Saji, H, Yamamoto, Y, Yamamoto, N, Nishimura, Y, Takahashi, T, Kuzushima, K, Emi, N & Akatsuka, Y 2014, 'Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H', Gene Therapy, vol. 21, no. 6, pp. 575-584. https://doi.org/10.1038/gt.2014.30

Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. / Inaguma, Y.; Akahori, Y.; Murayama, Y.; Shiraishi, K.; Tsuzuki-Iba, S.; Endoh, A.; Tsujikawa, J.; Demachi-Okamura, A.; Hiramatsu, K.; Saji, H.; Yamamoto, Y.; Yamamoto, Naoki; Nishimura, Y.; Takahashi, T.; Kuzushima, K.; Emi, N.; Akatsuka, Yoshiki.

In: Gene Therapy, Vol. 21, No. 6, 01.01.2014, p. 575-584.

Research output: Contribution to journalArticle

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T1 - Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H

AU - Inaguma, Y.

AU - Akahori, Y.

AU - Murayama, Y.

AU - Shiraishi, K.

AU - Tsuzuki-Iba, S.

AU - Endoh, A.

AU - Tsujikawa, J.

AU - Demachi-Okamura, A.

AU - Hiramatsu, K.

AU - Saji, H.

AU - Yamamoto, Y.

AU - Yamamoto, Naoki

AU - Nishimura, Y.

AU - Takahashi, T.

AU - Kuzushima, K.

AU - Emi, N.

AU - Akatsuka, Yoshiki

PY - 2014/1/1

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N2 - The genetic transfer of T-cell receptors (TCRs) directed toward target antigens into T lymphocytes has been used to generate antitumor T cells efficiently without the need for the in vitro induction and expansion of T cells with cognate specificity. Alternatively, T cells have been gene-modified with a TCR-like antibody or chimeric antigen receptor (CAR). We show that immunization of HLA-A2 transgenic mice with tetramerized recombinant HLA-A2 incorporating HA-1 H minor histocompatibility antigen (mHag) peptides and β2- microglobulin (HA-1 H/HLA-A2) generate highly specific antibodies. One single-chain variable region moiety (scFv) antibody, #131, demonstrated high affinity (K D =14.9 nM) for the HA-1 H/HLA-A2 complex. Primary human T cells transduced with #131 scFV coupled to CD28 transmembrane and CD3ζ domains were stained with HA-1 H/HLA-A2 tetramers slightly more intensely than a cytotoxic T lymphocyte (CTL) clone specific for endogenously HLA-A2-and HA-1 H-positive cells. Although #131 scFv CAR-T cells required >100-fold higher antigen density to exert cytotoxicity compared with the cognate CTL clone, they could produce inflammatory cytokines against cells expressing HLA-A2 and HA-1 H transgenes. These data implicate that T cells with high-affinity antigen receptors reduce the ability to lyse targets with low-density peptide/MHC complexes (∼100 per cell), while they could respond at cytokine production level.

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